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Microfluidic methods for non-thermal nucleic acid manipulations

  • US 6,406,893 B1
  • Filed: 11/20/2000
  • Issued: 06/18/2002
  • Est. Priority Date: 04/04/1997
  • Status: Expired due to Term
First Claim
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1. A method of amplifying a target nucleic acid by a non-thermal polymerase chain reaction, the method comprising:

  • providing a microfluidic device which includes a reaction chamber or channel, one or more sources of a target nucleic acid and primer sequences, a source of a chemical denaturant and a source of polymerase enzyme fluidly connected to the reaction chamber or channel, and a fluid direction system for delivering the chemical denaturant or the polymerase enzyme to the reaction chamber or channel;

    flowing the target nucleic acid and the primer sequences along the reaction chamber or channel;

    melting complementary strands of the flowing target nucleic acid by delivering a volume of the chemical denaturant to the reaction chamber or channel;

    annealing the primer sequences to the flowing target nucleic acid by eliminating a denaturing effect of the chemical denaturant;

    extending the primer sequences along the flowing target nucleic acid by delivering a volume of the polymerase enzyme to the reaction chamber or channel; and

    repeating the steps of melting, annealing and extending to amplify the flowing target nucleic acid.

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